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A flow-cytometry-based protocol for detection of mitochondrial ROS production under hypoxia
Hypoxia is known to stimulate mitochondrial reactive oxygen species (mROS) in cells. Here, we present a detailed protocol to detect mROS using MitoSOX staining in live cells under normoxia and hypoxia. Flow cytometry allows sensitive and reliable quantification of mROS by FlowJo software. We optimiz...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8086139/ https://www.ncbi.nlm.nih.gov/pubmed/33997804 http://dx.doi.org/10.1016/j.xpro.2021.100466 |
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author | Yang, Yun Zhang, Guimin Yang, Tao Gan, Jia Xu, Lin Yang, Hanshuo |
author_facet | Yang, Yun Zhang, Guimin Yang, Tao Gan, Jia Xu, Lin Yang, Hanshuo |
author_sort | Yang, Yun |
collection | PubMed |
description | Hypoxia is known to stimulate mitochondrial reactive oxygen species (mROS) in cells. Here, we present a detailed protocol to detect mROS using MitoSOX staining in live cells under normoxia and hypoxia. Flow cytometry allows sensitive and reliable quantification of mROS by FlowJo software. We optimized several aspects of the procedure including hypoxic treatment, working concentrations of the staining buffer, and quantitative analyses. Here, we use HepG2 cells, but the protocol can be applied to other cell lines. For complete details on the use and execution of this protocol, please refer to Yang et al. (2020). |
format | Online Article Text |
id | pubmed-8086139 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-80861392021-05-13 A flow-cytometry-based protocol for detection of mitochondrial ROS production under hypoxia Yang, Yun Zhang, Guimin Yang, Tao Gan, Jia Xu, Lin Yang, Hanshuo STAR Protoc Protocol Hypoxia is known to stimulate mitochondrial reactive oxygen species (mROS) in cells. Here, we present a detailed protocol to detect mROS using MitoSOX staining in live cells under normoxia and hypoxia. Flow cytometry allows sensitive and reliable quantification of mROS by FlowJo software. We optimized several aspects of the procedure including hypoxic treatment, working concentrations of the staining buffer, and quantitative analyses. Here, we use HepG2 cells, but the protocol can be applied to other cell lines. For complete details on the use and execution of this protocol, please refer to Yang et al. (2020). Elsevier 2021-04-20 /pmc/articles/PMC8086139/ /pubmed/33997804 http://dx.doi.org/10.1016/j.xpro.2021.100466 Text en © 2021 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Protocol Yang, Yun Zhang, Guimin Yang, Tao Gan, Jia Xu, Lin Yang, Hanshuo A flow-cytometry-based protocol for detection of mitochondrial ROS production under hypoxia |
title | A flow-cytometry-based protocol for detection of mitochondrial ROS production under hypoxia |
title_full | A flow-cytometry-based protocol for detection of mitochondrial ROS production under hypoxia |
title_fullStr | A flow-cytometry-based protocol for detection of mitochondrial ROS production under hypoxia |
title_full_unstemmed | A flow-cytometry-based protocol for detection of mitochondrial ROS production under hypoxia |
title_short | A flow-cytometry-based protocol for detection of mitochondrial ROS production under hypoxia |
title_sort | flow-cytometry-based protocol for detection of mitochondrial ros production under hypoxia |
topic | Protocol |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8086139/ https://www.ncbi.nlm.nih.gov/pubmed/33997804 http://dx.doi.org/10.1016/j.xpro.2021.100466 |
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